dual scRNA-seq analysis of P. vivax infected hepatocytes

Malaria-causing P. vivax parasites can linger in the human liver for weeks to months and then reactivate to cause recurrent blood-stage infection. While recognized as an important point of intervention for malaria eradication, very little is known about the molecular composition of the actively-replicating and quiescent intracellular P. vivax parasite populations, nor the molecular features and dynamics that underlie host cell responses to infection in humans. Here, we leverage a bioengineered human microliver platform to culture Thai clinical isolates of P. vivax in primary human hepatocytes and to conduct transcriptional profiling of infected cultures. By coupling targeted enrichment strategies with bulk and single-cell analyses, we capture both parasite and host transcripts in individual infected hepatocytes throughout the course of infection. We define stage-specific transcriptional signatures and identify a novel population of actively replicating parasites, sharing features with sexual blood-stages. Within infected liver cells, we find that infection suppresses transcription of key hepatocyte function genes, and that P. vivax elicits an innate immune response that can be manipulated to control infection in curative therapy mode. Our work provides an extendible framework and a valuable resource for understanding host-parasite interactions and reveals new insights into the biology of relapsing human malaria.致疟性间日疟原虫（P. vivax）可在人体肝脏内潜伏数周至数月，随后重新激活并引发复发性血液阶段感染。尽管该阶段被视为疟疾根除的关键干预靶点，但目前对活跃复制与休眠的细胞内间日疟原虫种群的分子组成，以及人类宿主细胞对感染产生应答的分子特征与调控动态，仍知之甚少。本研究借助生物工程化人类微型肝脏平台，在原代人肝细胞中培养泰国临床分离的间日疟原虫毒株，并对受感染的培养物开展转录组分析。通过将靶向富集策略与整体转录组及单细胞分析相结合，我们在感染全过程中，从单个受感染肝细胞中同时捕获疟原虫与宿主的转录本。本研究明确了阶段特异性转录特征，并鉴定出一类新型活跃复制的疟原虫种群，其特征与有性血液阶段疟原虫相似。在受感染的肝细胞中，我们发现感染会抑制肝细胞关键功能基因的转录，且间日疟原虫会触发先天免疫应答，该应答可通过治疗性干预手段调控以控制感染。本研究为解析宿主-疟原虫互作机制提供了可拓展的研究框架与宝贵的资源，并为复发性人类疟疾的生物学特性研究带来了新的见解。